Source: UNIV OF IDAHO submitted to
INHIBITION OF LISTERIA MONOCYTOGENES ON COLD-SMOKED SALMON BY EDIBLE ANTIMICROBIAL PACKAGING
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
0228666
Grant No.
(N/A)
Project No.
IDA01472
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Jul 1, 2012
Project End Date
Jun 30, 2017
Grant Year
(N/A)
Project Director
Unlu, GU.
Recipient Organization
UNIV OF IDAHO
875 PERIMETER DRIVE
MOSCOW,ID 83844-9803
Performing Department
School of Food Science
Non Technical Summary
Every year within the US, 76 million foodborne illnesses are reported with 325,000 hospitalizations and 5000 deaths. Costs associated with foodborne illnesses are approximately $23 billion every year. Over the past 30 years, listeriosis caused by L. monocytogenes has become a major foodborne disease. There are now over 1500 cases of listeriosis reported every year with 260 fatalities. Pregnant women, infants, and people with compromised immune system are at serious risk. Outbreaks of listeriosis in the early 1980s made the FDA and USDA establish a zero policy on Ready-to-eat (RTE) foods in which the occurrence of L. monocytogenes is considered unacceptable. Seafood has often been associated with foodborne listeriosis outbreaks. Occurrence of L. monocytogenes in RTE seafood ranges from 6% to 36% and can even reach 78%. The FDA/USDA risk assessment included smoked seafood in the list of foods that are at high risk of causing human listeriosis on a per annum basis. Consumer demand for minimally processed seafood has been on the rise. Therefore, there has been a great interest in developing efficient methods to control foodborne pathogens in foods by the use of naturally occurring antimicrobial agents. An alternative approach to control L. monocytogenes is the use of edible films with incorporated antimicrobial systems. Antimicrobial edible films can be used to improve microbiological food safety by reducing the risk of microbiological contamination by prolonged antimicrobial activity on the surface of the food. Edible films are made from various film forming substances that contain protein and carbohydrates. One of the main factors in choosing a raw material for film formation is the cost. The material chosen for packaging should not add much cost to the product. Potato processing waste is an inexpensive and readily available material. Each year approximately 10 billion pounds of potato processing waste is generated within the United States. Essential oils are volatile aromatic liquids obtained from various plant materials. Use of essential oils to preserve foods can ensure food safety by satisfying the consumers demand for minimally processed or natural foods. Essential oils from oregano, bay laurel, Spanish lavender and fennel show antibacterial activity against various foodborne pathogens. Of the essential oils studied, oregano essential oil has shown more antimicrobial activity against L. monocytogenes. The objectives of the study is to develop potato peel waste-based edible films with incorporated oregano essential oil, determine the physical properties of the films, and study their antimicrobial activity against L. monocytogenes inoculated onto the surface of cold-smoked salmon. The films have the potential to inhibit L. monocytogenes on cold-smoked salmon and other RTE seafood and thus ensure the safety of the foods. Demonstration of the effectiveness of the films developed in this study will likely increase the use of potato peel waste.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
7124010110025%
7124010202025%
7125010110025%
7125010202025%
Goals / Objectives
Our long-range goal is to develop edible films that can successfully be used in bioactive packaging of food products. The objective of this research proposal is to develop potato peel waste-based edible antimicrobial films and use them against Listeria monocytogenes. The specific objectives of the proposed research include: 1. Develop edible antimicrobial films from potato peel waste by incorporating oregano essential oil; 2. Evaluate the physical properties of the films; and 3. Evaluate the antimicrobial activity of the films against L. monocytogenes inoculated onto the surface of cold-smoked salmon.
Project Methods
Specific Objective 1: Develop edible antimicrobial films from potato peel waste by incorporating oregano essential oil. Edible films will be prepared from potato peel waste (PPW) using a method developed by Kang et al. (2010). Oregano essential oil will be incorporated into films at various concentrations. Specific Objective 2: Evaluate the physical properties of the films. Film thickness will be measured using Micrometer IP54. Water vapor permeability (WVP) will be measured by Gravimetric Modified Cup method that is based on ASTM standard method E 96-92.Tensile properties will be measured using the ASTM standard method D 882-01 and will include tensile strength (TS), elastic modulus (EM) (Young's modulus) and % elongation at break (%E). Pair wise comparisons equivalent to Fishers LSD will be conducted using Statistical Analysis Software (SAS) version 9.2. Specific Objective 3: Evaluate the antimicrobial activity of the films against L. monocytogenes inoculated onto the surface of cold-smoked salmon. Four strains of L. monocytogenes will be used in this study: V7 1/2a, CWD 1157, CWD 1198 and CWD1002. Minimum Inhibitory Concentration (MIC) of oregano essential oil will be determined by an agar well diffusion method modified from that of Winkowski et al. (1992). Antimicrobial activity of potato peel waste films with incorporated oregano essential oil (PPW-OO films) will be evaluated using microbiological growth medium and a method developed by Zivanovic et al. (2004). Vacuum packaged cold-smoked salmon (VP-CSS) with no added preservatives will be challenged with L. monocytogenes V7 1/2a. Growth of L. monocytogenes will be monitored in the presence of PPW-OO films for 28 days at 4oC. Inoculation will be done in two ways: (1) inoculating the surface of salmon with L. monocytogenes and then coating the surface with the PPW-OO film; (2) coating the surface of salmon with PPW-OO film and then inoculating L. monocytogenes on film surface. Inoculated and uncoated salmon as well as un-inoculated and uncoated salmon samples will serve as positive and negative controls. Experimental samples and controls will be vacuum-sealed and stored at 4oC for 28 days. Sampling will be done once a week for 4 weeks. At each sampling time point, samples will be stomached for 2 minutes in 0.1% peptone water and the resulting homogenates will be serially diluted and plated onto TAL plates. The TAL plates are double-layered plates with selective medium on the bottom and non-selective medium on the top (Kang et al., 1999). Oxford Medium will be used as the selective medium in bottom layer while non-selective Tryptic Soy Agar will be used as the top layer when preparing TAL plates. Colonies on TAL plates will be enumerated after incubation for 48 hours at 37oC. For storage studies, pooled ANOVA will be conducted assuming completely random design. Mean comparisons using Pair wise comparison will be conducted using SAS program. Two independent experiments will be conducted with each mean plus (or minus) standard deviation being the average of 4 replicates (two samples from each experiment).

Progress 07/01/12 to 06/30/17

Outputs
Target Audience:We have continued to reach scientific community and the food industry through our publications in the Journal of International Food Science and Technology , Journal of Food Science, and Journal of Food Chemistry and Nutrition. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? We have accomplished all three specific objectives as dicussed in former progress reports.

Publications


    Progress 10/01/15 to 09/30/16

    Outputs
    Target Audience:We have continued to reach scientific community and the food industry through our publications in the Journal of International Food Science and Technology (1), Journal of Food Science (1), and Journal of Food Chemistry and Nutrition (2). Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?We plan to complete our final report.

    Impacts
    What was accomplished under these goals? We have accomplished all three specific objectives as dicussed in former progress reports.

    Publications


      Progress 10/01/14 to 09/30/15

      Outputs
      Target Audience:We have reached the scientific community and the industry with interest in the area of biopackaging. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?A graduate student carried out various experiments for the project. Due in part to his efforts, the studentis well trained in the areas of bioactive food packaging. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?We plan to continue our work on inhibition of Listeria monocytogenes via the means of biopreservation.

      Impacts
      What was accomplished under these goals? Fish gelatin provides an alternative source of gelatin for Halal and Kosher applications and is prion and zoonotic agent free. However, applications of fish gelatin have been limited due to inferior mechanical and barrier properties. The physical properties of fish gelatins can be improved by crosslinking using natural polyphenolic compounds such as tannic acid. The objectives of this study were to develop fish gelatin films incorporated with tannic acid and evaluate their antioxidant, thermal, tensile, water vapor permeability and water solubility properties. Also, the effect of tannin-incorporated gelatins on the oxidative stability of salmon fillets was examined at 4 and 10 °C. Comparative data with bovine gelatins were generated. Fish gelatin (6.75% wt/wt) films were prepared at a gelatin:tannic acid ratio (wt/wt) of 1:0.05, 1:0.10 and 1:0.15. Tensile strength of bovine gelatin control was approximately 49 MPa and that of fish gelatin was 21 MPa. Tensile strength of bovine gelatins increased with tannic acid incorporation (P > 0.05) and did not vary significantly for fish gelatin films. Percent elongation of films increased and elastic modulus decreased with tannic acid incorporation. Water solubility of bovine gelatin films was reduced significantly (P < 0.05) and there was no effect of tannin on the solubility of fish gelatin films. The water vapor permeability was not significantly different for both the gelatins (P > 0.05) and the values ranged between 1.62 and 2.01 g mm/kPa h m2. Bovine and fish gelatin films with highest level of tannic acid showed an increase in glass transition temperature of approximately 12 and 6 °C, respectively. Films with tannic acid possess antioxidant activity and were able to reduce oxidation (TBARS values) in gelatin coated refrigerated salmon held for 12 days.

      Publications


        Progress 10/01/13 to 09/30/14

        Outputs
        Target Audience: We have reached the scientific community, specifically the audience of Journal of Food Chemistry and Nutrition, and the industry with interest in the areas of bioactive packaging. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? A Ph.D. recipient ('14) carried out various experiments for the project. Due in part to his research efforts, the professional is well trained in the areas of bioactive food packaging and microbial food safety. How have the results been disseminated to communities of interest? A manuscript was published. What do you plan to do during the next reporting period to accomplish the goals? Our research group plans to work on a review article and/or a book chapter on bioactive packaging.

        Impacts
        What was accomplished under these goals? Fish gelatin provides an alternative source of gelatin for Halal and Kosher applications and is prion and zoonotic agent free. However, applications of fish gelatin have been limited due to inferior mechanical and barrier properties. The physical properties of fish gelatins can be improved by crosslinking using natural polyphenolic compounds such as tannic acid. The objectives of this study were to develop fish gelatin films incorporated with tannic acid and evaluate their antioxidant, thermal, tensile, water vapor permeability and water solubility properties. Also, the effect of tannin-incorporated gelatins on the oxidative stability of salmon fillets was examined at 4 and 10 °C. Comparative data with bovine gelatins were generated. Fish gelatin (6.75% wt/wt) films were prepared at a gelatin:tannic acid ratio (wt/wt) of 1:0.05, 1:0.10 and 1:0.15. Tensile strength of bovine gelatin control was approximately 49 MPa and that of fish gelatin was 21 MPa. Tensile strength of bovine gelatins increased with tannic acid incorporation (P> 0.05) and did not vary significantly for fish gelatin films. Percent elongation of films increased and elastic modulus decreased with tannic acid incorporation. Water solubility of bovine gelatin films was reduced significantly (P< 0.05) and there was no effect of tannin on the solubility of fish gelatin films. The water vapor permeability was not significantly different for both the gelatins (P> 0.05) and the values ranged between 1.62 and 2.01 g mm/kPa h m2. Bovine and fish gelatin films with highest level of tannic acid showed an increase in glass transition temperature of approximately 12 and 6 °C, respectively. Films with tannic acid possess antioxidant activity and were able to reduce oxidation (TBARS values) in gelatin coated refrigerated salmon held for 12 days.

        Publications

        • Type: Journal Articles Status: Published Year Published: 2014 Citation: Tammineni, N., C. Nindo, B. Rasco, J. Powers, and G. �nl�. 2014. Tannin incorporated bovine and fish gelatin coatings: Effect on physical properties and oxidative stability of salmon fillets. Journal of Food Chemistry and Nutrition. 2 (2): 93-102.
        • Type: Theses/Dissertations Status: Published Year Published: 2014 Citation: Tammineni, N. 2014. Development of Fish Gelatin Films with Enhanced Physical, Antioxidant and Antimicrobial Properties. Ph.D. Dissertation. University of Idaho. Moscow, ID.


        Progress 01/01/13 to 09/30/13

        Outputs
        Target Audience: We have reached the scientific community and the industry with interest in the areas of bioactive packaging and microbial safety of seafood. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? A Ph.D. candidate and a MS student carried out various experiements for the project. Due in part to their efforts, the students are well trained in the areas of bioactive food packaging and microbial food safety. How have the results been disseminated to communities of interest? A manuscript was published. An abstract was published and the corresponding paper was presented at a scientific meeting. What do you plan to do during the next reporting period to accomplish the goals? We plan to continue our work on inhibition of Listeria monocytogenes on cold-smoked salmon and other seafood by edible antimicrobial packaging. Specifically, our research group plans to work on fish gelatin films crossed linked with tannic acid and evaluate their thermal, tensile and water vapor permeability properties.

        Impacts
        What was accomplished under these goals? In this study, potato peel waste (PPW)-based edible films with oregano oil were developed. Incorporation of oil into the films reduced the film strength and increased their water vapor pressure. The potato peel waste-oregano essential oil (PPW-OO) film reduced the growth of L. monocytogenes on cold-smoked salmon during storage under vacuum conditions at 4 °C for 28 days. The application of the newly developed PPW-OO films would provide an additional hurdle to the growth of L. monocytogenes on the surface of cold-smoked salmon. The PPW-OO films have the potential to be used as coatings on seafood and possibly other food products to inhibit L. monocytogenes. Our goal in a parallel study was to develop nisin-containing edible films using trout skin gelatin and test their effectiveness against L. monocytogenes on raw trout fillets (stored at 4 °C and 10 °C) as a strategy for reducing the risk of contamination. Films with 18 μg nisin/cm2 showed consistent inhibition and were chosen for storage studies conducted at 4 and 10 °C for 30 days. Trout fillets were challenged with 2 log CFU L. monocytogenes/g before or after coating with nisin-containing films. Films with nisin reduced L. monocytogenes counts below the detection limit (0.3 log CFU/g) at 4 °C. At 10 °C, a 0.3 to 1.1 log reduction was observed compared to controls by the end of storage. The effectiveness of treatments depended upon the concentration of nisin and storage temperature. The developed edible films have the potential to reduce pathogens on seafood and can be incorporated with a variety of antimicrobials.

        Publications

        • Type: Journal Articles Status: Published Year Published: 2013 Citation: Han, Yoo Tak, N. Tammineni, G. �nl�, B. Rasco, and C. Nindo. 2013. Inhibition of Listeria monocytogenes on rainbow trout (Oncorhynchus mykiss) using trout skin gelatin edible films containing nisin. Journal of Food Chemistry and Nutrition. ##plugins.citationFormat.abnt.location##, 1, jun. 2013. Available at: .
        • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Tammineni, N., G. �nl�, J. Powers, S. Sablani, B. Rasco, and C. Nindo. 2013. Properties of fish gelatin films crosslinked with tannic acid. Institute of Food Technologists (IFT) Annual Meeting 2013. Abstract 034-18. Chicago, IL.


        Progress 01/01/12 to 12/31/12

        Outputs
        OUTPUTS: Experiments were conducted. The results from the experiments were analyzed. A manuscript entitled "Development of antimicrobial potato peel waste-based edible films with oregano essential oil to inhibit Listeria monocytogenes on cold-smoked salmon was published in Journal of International Food Science and Technology to disseminate the research findings. PARTICIPANTS: Nothing significant to report during this reporting period. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

        Impacts
        Listeriosis caused by Listeria monocytogenes is a major foodborne illness. Foodborne listeriosis has been linked to the consumption of smoked salmon. Smoked salmon is a ready-to-eat product that is consumed without cooking. However, its pH, salt concentration and aw are within the permissible range for the growth of L. monocytogenes. Edible antimicrobial films allow controlled diffusion of antimicrobials to the surface of food, ensuring controlled release of antimicrobial activity. One of the objectives of our work was to develop edible antimicrobial films using potato peel waste, a new matrix, and oregano essential oil and study their effectiveness against L. monocytogenes on cold-smoked salmon. In our work, potato peel waste-based edible films with oregano oil (PPW-OO) were developed. Incorporation of oil into the films reduced the film strength and increased their water vapor permeability (WVP). The PPW-OO film reduced the growth of L. monocytogenes on cold-smoked salmon during storage under vacuum conditions at 4 degrees Celcius for 28 days. The application of the newly developed PPW-OO films would provide an additional hurdle to the growth of L. monocytogenes on the surface of cold-smoked salmon. The PPW-OO films have the potential to be used as coatings on seafood and possibly other food products to inhibit L. monocytogenes. Demonstration of the effectiveness of the PPW-OO film in this research will likely lead to increase in the use of the potato peel waste, a low-cost or no cost waste-stream.

        Publications

        • Tammineni, N., G. Unlu, and S. Min. 2012. Development of antimicrobial potato peel waste-based edible films with oregano essential oil to inhibit Listeria monocytogenes on cold-smoked salmon. Journal of International Food Science and Technology. Article first published online: 22 AUG 2012. DOI: 10.1111/j.1365-2621.2012.03156.x